Initiation of proximal-distal patterning in the vertebrate limb by signals and growth

Two broad classes of models have been proposed to explain the patterning of the proximal-distal axis of the vertebrate limb (from the shoulder to the digit tips). Differentiating between them, we demonstrate that early limb mesenchyme in the chick is initially maintained in a state capable of generating all limb segments through exposure to a combination of proximal and distal signals. As the limb bud grows, the proximal limb is established through continued exposure to flank-derived signal(s), whereas the developmental program determining the medial and distal segments is initiated in domains that grow beyond proximal influence. In addition, the system we have developed, combining in vitro and in vivo culture, opens the door to a new level of analysis of patterning mechanisms in the limb.     

鸡多趾性状研究进展

鸡常见趾型为四趾,北京油鸡等一些品种表现多趾特征.已有杂交试验证明鸡多趾性状为常染色体显性遗传但外显率不同.遗传图谱和比较基因组学分析显示,鸡2号染色体短臂端45cM处是鸡多趾性状关键候选区间.本文在参考前人工作基础上结合自己的研究工作,就鸡多趾性状表现类型、遗传规律、基因定位和候选基因等研究现状进行了综述.     

Diffusible signals, not autonomous mechanisms, determine the main proximodistal limb subdivision

Vertebrate limbs develop three main proximodistal (PD) segments (upper arm, forearm, and hand) in a proximal-to-distal sequence. Despite extensive research into limb development, whether PD specification occurs through nonautonomous or autonomous mechanisms is not resolved. Heterotopic transplantation of intact and recombinant chicken limb buds identifies signals in the embryo trunk that proximalize distal limb cells to generate a complete PD axis. In these transplants, retinoic acid induces proximalization, which is counteracted by fibroblast growth factors from the distal limb bud; these related actions suggest that the first limb-bud PD regionalization results from the balance between proximal and distal signals. The plasticity of limb progenitor cell identity in response to diffusible signals provides a unifying view of PD patterning during vertebrate limb development and regeneration.     

Embryological evidence identifies wing digits in birds as digits 1, 2, and 3

The identities of the digits of the avian forelimb are disputed. Whereas paleontological findings support the position that the digits correspond to digits one, two, and three, embryological evidence points to digit two, three, and four identities. By using transplantation and cell-labeling experiments, we found that the posteriormost digit in the wing does not correspond to digit four in the hindlimb; its progenitor segregates early from the zone of polarizing activity, placing it in the domain of digit three specification. We suggest that an avian-specific shift uncouples the digit anlagen from the molecular mechanisms that pattern them, resulting in the imposition of digit one, two, and three identities on the second, third, and fourth anlagens.     

Isolation and Analysis of α-Gliadin Gene Coding Sequences from Triticum durum

Three coding sequences of gliadins genes, designed as Gli2_Du1, Gli2_Du2 and Gli2_Du3, were isolated from the genomic DNA of Triticum durum accessions CItr5083. Gli2_Du1 and Gli2_Du2 contain 945 and 864 bp, encoding the mature proteins with 314 and 287 amino acid residues, respectively. Gli2_Du3 is recognized as a pseudogene due to the stop codon occurring in the coding region. The pseudogenes, commonly occurring in gliadins family, are attributed to the single base change C → T. The amino acid sequences deduced from these gene sequences were characterized with the typical structure of α-gliadin proteins, including the toxic sequences (PSQQQP). The peptide fraction PF(Y)PP(Q)is thought to be an extra unit of repetitive domain, slightly diverging from the previous report. Six cysteine residues were observed within two unique domains. Phylogenetic analysis showed Gli2_Du2 and Gli2_Du3 were closely related to the genes on chromosome 6A, whereas Gli2_Du1 seems to be more homologous with the genes on chromosome 6B.     

Isolation and Analysis of α-Gliadin Gene Coding Sequences from Triticum durum

Three coding sequences of gliadins genes, designed as Gli2_Dul, Gli2_Du2 and Gli2_Du3, were isolated from the genomic DNA of Triticum durum accessions CItr5083. Gli2_Dul and Gli2_Du2 contain 945 and 864 bp, encoding the mature proteins with 314 and 287 amino acid residues, respectively. Gli2_Du3 is recognized as a pseudogene due to the stop codon occurring in the coding region. The pseudogenes, commonly occurring in gliadins family, are attributed to the single base change C→T. The amino acid sequences deduced from these gene sequences were characterized with the typical structure of α-gliadin proteins, including the toxic sequences （PSQQQP）. The peptide fraction PF（Y）PP（Q）is thought to be an extra unit of repetitive domain, slightly diverging from the previous report. Six cysteine residues were observed within two unique domains. Phylogenetic analysis showed Gli2_Du2 and Gli2_Du3 were closely related to the genes on chromosome 6A, whereas Gli2_Dul seems to be more homologous with the genes on chromosome 6B.     

‘红地球’葡萄花芽分化过程中的转录组分析

【目的】 葡萄是我国重要的果树树种,花芽分化直接影响葡萄的质量和数量。对‘红地球’葡萄花芽分化过程中的花芽进行比较分析,探索‘红地球’葡萄花芽分化机制,挖掘关键基因,为了解‘红地球’葡萄花芽分化提供理论基础。【方法】 对‘红地球’葡萄花芽分化过程中4个发育阶段：S1（未分化期）、S2（花原始体发育期）、S3（花序主轴发育期）和S4（花序二级轴发育期）的芽进行形态学观察和植物激素测定,并进行转录组测序分析及验证。【结果】 ‘红地球’葡萄花芽分化过程中共发现13 729个差异基因,其中S1-S2、S2-S3、S3-S4和S1-S4分别有4 158、2 050、3 425和7 652个差异基因。在S1-S4差异基因的富集调控网络中发现差异基因在激素介导的信号通路、脱落酸代谢过程、对酸性化学物质的反应和植物细胞壁组织或生物发生等通路富集。在激素介导的信号通路中发现大量与生长素、赤霉素和脱落酸等相关基因,测定表明,生长素在S2时期含量最高,而在S3和S4时期含量最低;赤霉素含量在花芽分化过程中不断降低,在S4时期为S1时期的80%;脱落酸含量在S1和S4时期较高,而在S2时期最低。此外,S1-S4差异基因来自转录因子家族（MYB、ERF、bHLH和MADS-box等）,表明这些转录因子家族基因参与了‘红地球’葡萄花芽分化。对差异表达的13个MADS-box家族基因进一步分析表明,MADS8、AGL65、AGL15、AGL12和MADS2在花芽分化进程中表达上调,而AGL30、LeMADS、FBP24、AGL14和MADS3表达下调。对这些MADS-box基因进行qRT-PCR验证,基因表达趋势与转录组数据一致且相关系数较高,表明数据分析结果可靠。【结论】 ‘红地球’葡萄花芽分化是一个复杂的生物过程,其中,植物激素介导的信号通路以及MADS-box家族基因在花芽分化中发挥重要作用。研究结果提供了一个关于转录因子、基因和激素的信息,有助于揭开这一复杂的发育过程,并为‘红地球’葡萄花芽分化综合模型的建立提供理论基础。     

黄牛Gli3基因第10外显子多态性及其与生长性状相关性

利用PCR-SSCP和DNA测序技术研究708头黄牛Gli3基因第10外显子的多态性,发现此外显子存在SSCP多态。对不同SSCP带型的对应片段进行测序分析,共发现2个新的SNP多态位点(NW_001494928:g.205649TC,205754AC),这2个SNP完全连锁。南阳牛不同基因型与生长发育性状的相关性分析显示,CC基因型个体的24月龄平均日增体质量指标显著大于TT基因型个体(P0.05),其余各项指标基因型间差异不显著(P0.05)。因此,该位点有可能作为南阳牛生长性状辅助选择的标记之一。秦川牛和郏县红牛群体中没有发现与该基因座多态显著相关的性状。     

鲁枣1号组织培养和植株再生体系研究

以鲁枣1号为试材,以正在生长的枣头嫩枝为外植体,进行了芽的诱导和试管苗快繁研究。结果表明,嫩枝茎段上未萌发的主芽在启动培养基(MS+1 mg/L BA+0.2 mg/L IBA+3%蔗糖)上可萌发成新梢,萌发率为54.8%。在相同的启动培养基上,远离培养基的茎段切口处可诱导产生不定芽,不定芽再生率为23.8%。不定芽和主芽新梢在MS+2 mg/L BA+0.4 mg/L IAA+3%蔗糖+6 g/L琼脂培养基上增殖生长良好。试管绿苗在1/4MS+0.5 mg/L IBA+2%蔗糖+6 g/L琼脂培养基上生根率达70%以上。     

小麦品种“川农16”α-醇溶蛋白基因序列分析

 【目的】克隆和分析“川农16”醇溶蛋白基因,为其进一步遗传改良提供更多依据。【方法】根据已报道的α-醇溶蛋白基因序列设计引物,对小麦品种“川农16”总DNA进行PCR扩增得到约900 bp的DNA片段,分离纯化后连接到pMD18-T载体上,转化后筛选阳性克隆进行测序。【结果】获得4个不同的基因序列：Gli2-CN16-9、Gli2-CN16-12、Gli2-CN16-14和Gli2-CN16-6,GenBank登录号分别为DQ246446、DQ246447、DQ246448和DQ246449。其中,Gli2-CN16-9、Gli2-CN16-12和Gli2-CN16-14分别为861、870和900 bp,可分别编码286、289和299个氨基酸残基的成熟蛋白;而Gli2-CN16-6编码区长度为852 bp,由于存在2个提前终止密码子,不能编码有功能的成熟蛋白,为假基因。【结论】序列比较显示它们与α-醇溶蛋白基因有很高的一致性;与γ-和ω-醇溶蛋白基因差异明显。     

柱穗山羊草α-醇溶蛋白基因的克隆与序列分析

根据已知的普通小麦α-醇溶蛋白基因序列设计引物,采用PCR方法克隆基因并进行序列分析。从柱穗山羊草Y127中克隆得到1个α-醇溶蛋白基因序列Gli2-Z-2,它具有α-醇溶蛋白基因的典型结构特征,编码区全长939 bp,编码313个氨基酸。氨基酸序列比较显示,Gli2-Z-2在多聚谷氨酰胺区比已报道的α-醇溶蛋白序列有较多的谷氨酰胺残基。     

南通地区国兰生长发育动态研究初报

[目的]研究南通地区国兰的生长发育动态,为国兰的栽培管理提供理论基础。[方法]以春兰和蕙兰主栽品种为试材,定期观测叶芽、花芽的生长发育情况。[结果]春兰春芽（叶芽）快速生长期在4月中旬至6月下旬,蕙兰春芽（叶芽）旺盛生长期在4月中旬至7月中旬;春兰秋芽（叶芽）快速生长期在10月初至12月中旬;蕙兰秋芽（叶芽）旺盛生长期在10月初至12月下旬。春兰、蕙兰的花葶均在10月底出土,但春兰的花葶在1月底进入缓慢生长期,而蕙兰的花葶则在2月底才进入缓慢生长期。[结论]南通地区春兰和蕙兰的主要施肥期宜为：春季3月下旬至4月初,以氮肥为主;秋季9月中下旬,宜多施磷、钾肥。     

The limb bud Shh-Fgf feedback loop is terminated by expansion of former ZPA cells

Vertebrate limb outgrowth is driven by a positive feedback loop involving Sonic Hedgehog (Shh), Gremlin, and Fgf4. By overexpressing individual components of the loop at a time after these genes are normally down-regulated in chicken embryos, we found that Shh no longer maintains Gremlin in the posterior limb. Shh-expressing cells and their descendants cannot express Gremlin. The proliferation of these descendants forms a barrier separating the Shh signal from Gremlin-expressing cells, which breaks down the Shh-Fgf4 loop and thereby affects limb size and provides a mechanism explaining regulative properties of the limb bud.     

施用6-BA对春石斛花芽分化及内源激素的影响

该文研究了施用6-BA对春石斛品种花芽分化及ZT、IAA、 ABA、 GA3的影响。结果表明:施用浓度为1 000 mg/L 6-BA,极显著地提高了春石斛花芽形成个数和单株花芽形成率,并促使花芽分化的进程提前;施用1 000 mg/L 6-BA提高了花芽内ZT的含量,降低了GA3和ABA的含量,花芽形态分化前IAA含量降低,而后IAA含量升高; 施用1 000 mg/L 6-BA提高了春石斛花芽内ZT/GA3、ZT/IAA、ZT/ABA的比值,降低了ABA/IAA的比值,这可能促进了花芽的分化和形成。      

非洲菊不定芽增殖的影响因素

以非洲菊不定芽为试材,研究了6-BA、NAA和CC组合以及GA3、蔗糖和琼脂组合对不定芽增殖和发育的影响。结果表明,6-BA 3mg/L、NAA 0.3 mg/L和CC 3 mg/L组合;GA30.5 mg/L、蔗糖40 g/L和琼脂6 g/L组合,增殖倍数最大,植株生长健壮,最适合非洲菊不定芽增殖。     

绿竹花发育相关基因BoAP3的克隆与分析

MADS—box基因家族在决定花分生组织特性和花器官发育过程中起着重要的作用。以绿竹Bambusaoldhamii开花试管苗花芽为植物材料,采用cDNA末端快速扩增技术（rapid amplification of cDNAends,RACE）技术,获得了1条MADS—box基因家族的基因,命名为BoAP3。序列分析结果表明：BoAP3开放阅读框（open reading frame,ORF）长度为654bp,编码218个氨基酸,具有典型的植物MADS—box蛋白结构,其编码肽链包含了MADS区、K区、I区和C区。B胡丹与小麦Triticum aestivum,水稻Oryzasatva等AP3-like同源基因所编码的氨基酸同源性达到80％以上。定量聚合酶链式反应（PCR）结果表明：BoAP3基因在开花试管苗的花芽中表达量是不开花试管苗营养芽表达量的8．1倍,表明该基因可能参与了花器官的发育。     

杂交中稻再生芽生长的影响因素研究

以18个杂交中稻组合为材料,研究了杂交中稻地上部植株性状和施氮对再生芽生长的影响。结果表明,头季稻齐穗期单茎干物质积累量越大、成熟期满足头季籽粒灌浆后茎鞘中剩余光合物质量越多,其再生芽活芽率与出苗率越高。高再生芽活芽率和出苗率的头季稻植株重要性状表现为株高偏矮、成穗率较高。在保持较高活芽率的基础上,提高后期根系活力是进一步提高再生稻产量的重要途径。